Matsubara Yumiko, Murata Mitsuru, Ikeda Yasuo
Department of Laboratory Medicine, School of Medicine, Keio University, Tokyo, Japan.
Methods Mol Biol. 2012;788:249-58. doi: 10.1007/978-1-61779-307-3_17.
The molecular mechanisms whereby stem cells develop into platelet-producing megakaryocytes (MKs) are not yet fully understood. Within this chapter we describe a two-step in vitro culture system in which MKs and platelets are generated from primary subcutaneous adipose tissues and the preadipocyte cell line 3T3L1. The cells are first cultured in an adipocyte induction medium for 10-12 days, followed by 8-14 days culture in a MK differentiation medium. Adipose tissue-derived MKs and platelets display a number of morphological and functional characteristics (e.g., secretory granules, open canalicular membranes) comparable with the native cell type. The use of subcutaneous adipose tissue to produce a large number of platelets is advantageous because this tissue is easily obtained and available in large quantities. Thus, this in vitro culture system may prove useful in both regenerative medicine, but it may also be used in understanding fundamental research questions within MK and platelet research, including further elucidation of the pathways that cause cells to differentiate along the MK lineage ultimately leading to platelet production.
干细胞发育成产生血小板的巨核细胞（MKs）的分子机制尚未完全明确。在本章中，我们描述了一种两步体外培养系统，其中MKs和血小板由原发性皮下脂肪组织和前脂肪细胞系3T3L1生成。细胞首先在脂肪细胞诱导培养基中培养10 - 12天，随后在MK分化培养基中培养8 - 14天。脂肪组织来源的MKs和血小板表现出许多与天然细胞类型相当的形态和功能特征（例如，分泌颗粒、开放小管膜）。利用皮下脂肪组织大量生产血小板具有优势，因为这种组织易于获取且数量丰富。因此，这种体外培养系统可能在再生医学中证明是有用的，但它也可用于理解MK和血小板研究中的基础研究问题，包括进一步阐明导致细胞沿MK谱系分化最终产生血小板的途径。
